Production of Cellulase by Trichoderma viride via Submerged Fermentation using Corncob as Substrate

Kamaluddeen, S.Y., Madika, A.
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Abstract

Cellulolytic enzymes are commonly used in lignocellulosic biomass conversion, textile, and paper and pulp industries. However, the high cost of cellulase and the high demand has led to increase researches to explore agro-residues for cost-effective production. This study was aimed at isolating Trichoderma viride from decaying wood and soil, and testing the isolates for cellulase production via submerged fermentation using corncob as substrate. Twenty-one (21) soil samples were collected from three different locations (botanical garden, refuse dump site and decaying wood) and used in the study. Colonies suspected to be Trichoderma viride were identified by microscopy of lactophenol cotton blue-stained preparation and slide culture technique, respectively. Isolates confirmed to be Trichoderma viride were then screened for cellulase production on a mineral salt medium containing carboxymethyl cellulose as an inducer. The isolates were further tested for cellulase production using corncob as the substrate via submerged fermentation. Out of the 21 samples analysed, 4 (19%) were positive for Trichoderma viride based on cultural and microscopic characteristics. The highest occurrence of Trichoderma viride (57%) was recorded from decaying wood samples, while no occurrence (0%) was observed from the botanical garden and refuse dumpsite soils. All four isolates obtained produced cellulase, with T. viride DW6 having the highest zone of cellulose hydrolysis (61.5 ± 2.12 mm) and T. viride DW1 having the least (58.5 ± 2.12 mm). The highest enzyme activity was recorded from T. viride DW4 (3.41 ± 0.46 U/ml) and the lowest from T. viride DW1 (1.77 ± 0.52 U/ml) using corncob as substrate. Trichoderma viride is abundantly present in decaying wood, and such isolates have high cellulase-producing ability, as wood material is rich in cellulose. This preliminary study provides information on the isolation of T. viride from decaying wood and their potential for cellulase production using corncob as substrate.
以玉米芯为底物的绿木霉深层发酵生产纤维素酶的研究
纤维素水解酶通常用于木质纤维素生物质转化、纺织、造纸和纸浆工业。然而,纤维素酶的高成本和高需求导致人们越来越多地研究探索农业残留物的成本效益生产。本研究旨在从腐烂的木材和土壤中分离出绿色木霉,并以玉米芯为底物进行深层发酵生产纤维素酶的试验。从三个不同的地点(植物园、垃圾场和腐烂的木材)收集21个土壤样本用于研究。分别采用乳酚棉蓝染色镜检和玻片培养法对疑似绿色木霉菌落进行鉴定。然后在含有羧甲基纤维素作为诱诱剂的无机盐培养基上筛选证实为绿色木霉的分离株,以生产纤维素酶。以玉米芯为底物进行深层发酵,进一步测试了分离菌株生产纤维素酶的能力。在分析的21份样本中,根据培养和显微镜特征,4份(19%)呈绿色木霉阳性。腐木样中绿木霉的发生率最高(57%),而植物园和垃圾场土壤中没有发现绿木霉(0%)。4个菌株均能产生纤维素酶,其中绿色T. DW6的纤维素水解区最高(61.5±2.12 mm),而绿色T. DW1的纤维素水解区最低(58.5±2.12 mm)。以玉米芯为底物的绿霉DW4酶活性最高(3.41±0.46 U/ml),最低(1.77±0.52 U/ml)。绿色木霉大量存在于腐烂的木材中,由于木材中含有丰富的纤维素,这种菌株具有较高的纤维素酶生产能力。这项初步研究提供了从腐烂木材中分离出绿霉菌及其以玉米芯为底物生产纤维素酶的潜力的信息。
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